Nucleic acid sequence encoding ovarian antigen, CA125, and uses thereof

Abstract

The present invention provides an isolated nucleic acid molecule comprising sequences encoding the CA125 protein or a portion thereof. This invention also provides a method to detect ovarian cancer in a subject. Furthermore, this invention provides a method for the diagnosis of a cancer which expresses CA125 by detecting CA125-expressing cells in the blood or other fluids of patients. This invention also provides a method of producing CA125 protein. Finally, this invention provides a method to treat or prevent cancer using a vaccine comprising CA125 nucleic acid or protein.

What is claimed is:  
     
         1 . An isolated nucleic acid molecule comprising sequences encoding the CA125 protein or a portion thereof.  
     
     
         2 . The gene encoding the CA125 protein.  
     
     
         3 . The isolated nucleic acid molecule of  claim 1  comprising sequence set forth in FIG. 6 and the corresponding CA125 protein comprising sequence set forth in FIG. 5.  
     
     
         4 . The isolated nucleic acid molecule of  claim 1  comprising sequence set forth in FIG. 7 and the corresponding CA125 protein sequence set forth in FIG. 8.  
     
     
         5 . The nucleic acid of  claim 1  comprising sequence set forth in FIG. 11.  
     
     
         6 . The nucleic acid of  claim 1  encoding protein comprising at least a portion of the amino acid sequence set forth in FIG. 12.  
     
     
         7 . The gene of  claim 2  comprising sequence set forth in FIG. 10.  
     
     
         8 . The isolated nucleic acid molecules of  claim 1 ,  2 ,  3 ,  4 ,  5 ,  6 , or  7 , wherein the nucleic acid is RNA, cDNA, genomic DNA, or synthetic DNA.  
     
     
         9 . A vector comprising the nucleic acid molecule of  claim 1 ,  2 ,  3 ,  4 ,  5 ,  6 ,  7 , or  8 .  
     
     
         10 . The vector of  claim 9 , designated as pBK-CMV-B4 comprising sequence set forth in FIG. 6 and the corresponding CA125 protein comprising sequence set forth in FIG. 5.  
     
     
         11 . The vector of  claim 9 , designated as pBKCMV-B30 comprising sequence set forth in FIG. 7 and the corresponding CA125 protein comprising sequence set forth in FIG. 8.  
     
     
         12 . The vector of  claim 9 , designated as pCMV-Tag-B4 comprising sequence set forth in FIG. 6 and the corresponding CA125 protein comprising sequence set forth in FIG. 5.  
     
     
         13 . The vector of  claim 9 , designated as pCMV-Tag-B30 comprising sequence set forth in FIG. 7 and the corresponding CA125 protein comprising sequence set forth in FIG. 8.  
     
     
         14 . An expression system comprising the vector of  claim 9 .  
     
     
         15 . The expression system of  claim 14 , wherein the system is a eukaryotic or prokaryotic system.  
     
     
         16 . A method for producing CA125 protein comprising the expression system of  claim 14 .  
     
     
         17 . An isolated nucleic acid molecule comprising sequence capable of specifically hybridizing to the sequences of  claim 1  or  2 .  
     
     
         18 . The nucleic acid molecule of  claim 17  capable of inhibiting the expression of the CA125 protein.  
     
     
         19 . A method of inhibiting expression of CA125 inside a cell by vector-directed expression of an RNA able to hybridize with the RNA of CA125.  
     
     
         20 . The nucleic acid molecule of  claim 17  or  18  which is at least a 10 mer.  
     
     
         21 . The nucleic acid molecule of  claim 17  or  18  which is at least a 20 mer.  
     
     
         22 . A method to detect ovarian cancer in a subject comprising steps of: 
 a) contacting the isolated nucleic acid molecule of  claim 17  with RNA from a sample from the subject under conditions permitting the formation of a hybrid complex, and    b) detecting the hybrid complex, wherein a positive detection indicates the expression of the antigen and presence of cancer.    
     
     
         23 . A method of monitoring ovarian cancer therapy in a subject comprising steps of: 
 a) contacting the isolated nucleic acid molecule of  claim 17  with RNA from a sample from the subject under conditions permitting the formation of a hybrid complex, and    b) measuring the amount of the hybrid complex, wherein a decrease in the hybrid complex indicates the success of therapy.    
     
     
         24 . A method for inhibiting the expression of the CA125 protein comprising contacting an appropriate amount of the nucleic acid molecule of  claim 17  or  18  so that hybridization of the gene or transcript encoding the CA125 protein will occur, thereby inhibiting the expression of the protein.  
     
     
         25 . A composition comprising the isolated nucleic acid molecule of  claim 17  or  18 .  
     
     
         26 . A vaccine for a cancer which expresses CA125 protein comprising an appropriate amount of the isolated nucleic acid molecules of  claim 1  or  2 .  
     
     
         27 . A vaccine for a cancer which expresses CA125 protein comprising an appropriate amount of an expression vector with the nucleic acid molecules which, when expressed, are capable of producing a product which induces an immune response to CA125 protein.  
     
     
         28 . The vaccine of  claim 27 , wherein the nucleic acid molecule comprises sequences encoding human CA125 protein or a portion thereof.  
     
     
         29 . The vaccine of  claim 28 , wherein the expressed human sequence is linked to a carrier.  
     
     
         30 . The vaccine of  claim 27 , wherein the nucleic acid molecule comprises a nonhuman sequence.  
     
     
         31 . The vaccine of  claim 27 , wherein the nucleic acid molecule comprises a primate sequence.  
     
     
         32 . The vaccine of  claim 27 , wherein the nucleic acid molecule comprises a murine sequence.  
     
     
         33 . The vaccine of  claim 27 , wherein the nucleic acid molecule comprises a synthetic sequence, which, when expressed, is capable of producing a product which induces an immune response to CA125 protein.  
     
     
         34 . The vaccine of  claim 33 , wherein the sequence hybridizes with or is homologous to the sequences encoding human CA125 protein.  
     
     
         35 . The vaccine of claims  26 - 34 , further comprising a suitable adjuvant.  
     
     
         36 . The vaccine of claims  26 - 34 , wherein the adjuvant is an alum.  
     
     
         37 . The vaccine of claims  26 - 36 , wherein the cancer is an ovarian, pancreatic, breast, endometrial, or lung carcinoma.  
     
     
         38 . A method to treat a cancer which expresses CA125 in a subject comprising administering to the subject an appropriate amount of the vaccine of claims  26 - 36 .  
     
     
         39 . The method of  claim 38 , wherein the cancer is an ovarian, pancreatic, breast, endometrial, or lung carcinoma.  
     
     
         40 . A vaccine for a cancer which expresses CA125 comprising an appropriate amount of the expressed CA125 protein corresponding to the sequence in  claim 1 .  
     
     
         41 . A vaccine for a cancer which expresses CA125 protein comprising an appropriate amount of a substance which induces an immune response to CA125 protein.  
     
     
         42 . The vaccine of  claim 41 , wherein the substance is a polypeptide or a peptide.  
     
     
         43 . The vaccine of  claim 42 , wherein the polypeptide comprises sequences encoding human CA125 protein or a portion thereof.  
     
     
         44 . The vaccine of  claim 43 , wherein the expressed human sequence is linked to a carrier.  
     
     
         45 . The vaccine of  claim 41 , wherein the polypeptide comprises a nonhuman sequence.  
     
     
         46 . The vaccine of  claim 45 , wherein the polypeptide comprises a primate sequence.  
     
     
         47 . The vaccine of  claim 45 , wherein the polypeptide comprises a murine sequence.  
     
     
         48 . The vaccine of  claim 42 , wherein the polypeptide comprises a synthetic sequence, which, when expressed, is capable of producing a product which induces immune response to CA125 protein.  
     
     
         49 . The vaccine of claims  40 - 48 , further comprising a suitable adjuvant.  
     
     
         50 . The vaccine of  claim 49 , wherein the adjuvant is an alum.  
     
     
         51 . The vaccine of claims  40 - 50 , wherein the expressed protein is conjugated to a protein carrier to increase the immunogenicity.  
     
     
         52 . The vaccine of claims  40 - 51 , wherein the cancer is an ovarian, pancreatic, breast, endometrial, or lung carcinoma.  
     
     
         53 . A method to treat a cancer which expresses CA125 in a subject comprising administering to the subject an appropriate amount of the vaccine of claims  40 - 51 .  
     
     
         54 . A method to prevent a cancer which expresses CA125 in a subject comprising administering to the subject an appropriate amount of the vaccine of claims  40 - 51 .  
     
     
         55 . The method of claims  53  or  54 , wherein the cancer is an ovarian, pancreatic, breast, endometrial, or lung carcinoma.  
     
     
         56 . A method for the diagnosis of a cancer which expresses CA125 by detecting CA125-expressing cells in the blood or other fluids of patients based on the nucleic acid sequence which encodes CA125.  
     
     
         57 . A method for monitoring the therapy of a cancer which expresses CA125 by measuring the expression of CA125-expressing cells in the blood or other fluids of patients based on the nucleic acid sequence which encodes CA125, a decrease of either the number of CA125-expressing cells or level of protein expression in the cell, indicating the success of the therapy.  
     
     
         58 . The method of  claim 56  or  57 , wherein the detection is based on polymerase chain reaction with appropriate primers.  
     
     
         59 . A method of producing CA125 protein comprising steps of: 
 a) constructing a vector adapted for expression in a cell which comprises the regulatory elements necessary for expression of nucleic acid in the cell operatively linked to the nucleic acid encoding the CA125 protein so as to permit expression thereof;    b) placing the cells of step (a) under conditions allowing the expression of the CA125 protein; and    c) recovering the CA125 protein so expressed.    
     
     
         60 . The method of  claim 59 , wherein the cell type is selected from the group consisting of bacterial cells, yeast cells, insect cells, and mammalian cells.  
     
     
         61 . The CA125 protein expressed by the method in  claim 59  or  60 .  
     
     
         62 . A method for production of antibodies against CA125 protein using the protein of  claim 61 .  
     
     
         63 . Antibodies produced by the method of  claim 62 .  
     
     
         64 . A method for monitoring the therapy of cancer which expresses CA125 using the antibodies of  claim 63 .  
     
     
         65 . A method of diagnosis of cancer which expresses CA125 using the antibodies of  claim 63 .  
     
     
         66 . A method for determining the immunoreactive part of CA125 comprising contacting antibodies which are known to be reactive to CA125 with the protein of  claim 61 .  
     
     
         67 . A transgenic nonhuman organism comprising the isolated nucleic acid molecule of  claim 1  or  2 .  
     
     
         68 . A transgenic nonhuman mammal of  claim 67 .  
     
     
         69 . A nonhuman organism, wherein the expression of CA125 is inhibited.  
     
     
         70 . The nonhuman mammal of  claim 69 .  
     
     
         71 . The nonhuman mammal of  claim 70 , wherein the mammal is a mouse.  
     
     
         72 . A method for screening a compound for treatment of cancer which expresses CA125 protein comprising administering the compound to the transgenic nonhuman organism of claims  67 - 71 , a decrease in expression of CA125 protein indicating that the compound may be useful for treatment of the cancer.